Conjugation of nucleic acid probes to 6-aminoglucose‐based homo- and copolymers. II. Application to diagnostics

Water-Soluble Dendrigrafts Bearing Saccharidic Moieties: Elaboration and Application to Enzyme Linked OligoSorbent Assay (ELOSA) Diagnostic Tests

“…In comparison, the same test without dendrigrafts (with capture ODNs alone) reached the same detection limit for 10 11 DNA copies per mL and thus was 100 times less sensitive. Moreover, the background signal provided by the IV-ODN conjugate was relatively low (1000 RFU, a value similar to that of linear copolymers) (9).…”

“…1 List of abbreviations: DLS, dynamic light scattering; DMF, dimethyl formamide; DMSO, dimethyl sulfoxide; EGP, 1,2:3,4-di-Oisopropylidene-6-O-epoxypropyl-D-galactopyranose; ELISA, enzymelinked immunosorbent assay; ELOSA, enzyme-linked oligosorbent assay; DiGVE, 1,2:3,4-di-O-isopropylidene-6-O-(2-vinyloxy ethyl)-Dgalactopyranose; Gal, galactose; HBV, hepatitis B virus; NMP, N-methylpyrrolidone; ODN, oligonucleotide; PCEVE, poly(chloroethyl vinyl ether); PDiPGVE, poly ( Concerning polymer-ODN conjugate properties desirable for an effective enhancement of the signal, it was shown that accessibility of the immobilized ODNs (rather than their number) was a major issue. Among the above-mentioned functional (co)polymers, various architectures have been designed such as (i) linear (co)polymers, namely poly(maleic anhydride-co-methyl vinyl ether) (8), poly(N-vinylpyrrolidoneco-N-acryloxysuccinimide) (7,10), poly(N-(2,2-dimethoxyethyl)-N-methyl acrylamide) (11), and synthetic glycopolymers (9,12,13), (ii) graft (co)polymers (14,15), and (iii) latex particles (18,19). However, to our knowledge, no study describing the use of dendrigrafts as supports for ODN immobilization has been previously reported.…”

“…An alternative way consists of the preparation of (co)polymer−ODN amplification structures (conjugates). With this aim, a number of new polymer supports, consisting of functional (co)polymers bearing different reactive groups (activated ester, anhydride, aldehyde), have been developed to immobilize ODNs ( − ). The conjugates were typically prepared either by binding amino-ended single-stranded oligonucleotide probes (synthesized via automated synthesis with an −NH 2 group at the 5‘ position) onto reactive (co)polymers via amination or amidation reaction ( − ) or by direct synthesis of the ODNs from the polymer backbone ( − ).…”

“…With this aim, a number of new polymer supports, consisting of functional (co)polymers bearing different reactive groups (activated ester, anhydride, aldehyde), have been developed to immobilize ODNs ( − ). The conjugates were typically prepared either by binding amino-ended single-stranded oligonucleotide probes (synthesized via automated synthesis with an −NH 2 group at the 5‘ position) onto reactive (co)polymers via amination or amidation reaction ( − ) or by direct synthesis of the ODNs from the polymer backbone ( − ). Both kinds of conjugates enable the capture of complementary DNA targets through hybridization and lead to an increase in the sensitivity of the ELOSA test.…”

Nucleic Acid Analysis

“…In combination with reverse transcriptase-polymerase chain reaction, the assay could detect prostate-specific antigen mRNA from a single cancer cell amidst 1 million normal cells (38). Nucleic acid probes were conjugated to 6-aminoglucose homo-and copolymers and applied to the quantification of hepatitis B virus (39). During slot blot analysis of adeno-associated virus it was found that Mg 2+ inhibits the binding of viral DNA, released from disrupted virions, to nylon and nitrocellulose membranes (40).…”